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CHAPTER 1

THE PROBLEM AND ITS BACKGROUND

1.1 Introduction

Coagulation also known as blood clotting is the process by which the

blood changes from liquid to a gel, forming a blood clot that stop the blood

from the wounds. It potentially results in hemostasis, the cessation of blood loss

from a damage vessel, followed by repair (David Lollicrap, 2009). The

mechanism of coagulation involves activation, adhesion, and aggregation of

platelets along with deposition and maturation of fibrin.

Normal blood clotting depends on the presence in the blood of certain

proteins called clotting factors. Those people who lack one of these clotting

factors often due to an inherited disorder may be given specific blood products,

which are concentrated supplements of the missing protein. For example, a

blood product called Factor VIII is needed for the treatment of the inherited

bleeding disorder haemophilia, in which a defective gene causes a deficiency of

natural Factor VIII in the blood. Another blood product, fresh frozen plasma, is

given to counteract abnormally prolonged or severe bleeding due to causes such

as an excessive dose of anticoagulants (Medical Guide, 2010).

But other proven studies said that there are many herbal medicines that

promote blood coagulation such as these certain products containing stinging

nettle have shown, when applied to the skin, can reduce bleeding during

surgery. The product, called Ankaferd blood stopper, is made up of alpinia,

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licorice, thyme, common grape vine and stinging nettle, and has also shown

evidence of reducing bleeding after dental surgery. In Chinese researchers they

used Persicae Semen (Taoren) and Carthami Flos (Honghua) used in pair which

is named as Taoren–Honghua (TH) herb pair has been used in traditional

Chinese medicine (TCM) for promoting blood circulation to dissipate blood

stasis for many years in China. Other researches also said that a common

pathological syndrome in the elderly called blood stasis. In traditional Chinese

medicine, the treatment for the syndrome is by herbs which activates blood

circulation to remove blood stasis. The herbs have the efficacy of improving

hemorheological events. Therefore, the herbs are the source for developing

hemorheologicals. Ligustrazine isolated from Chuangxiong is an example. It

showed significant inhibition on shear induced platelet aggregation and on

platelet intracellular calcium demonstrated by laser confocal microscope.

In the article titled better health care it is stated that side effect can be

caused by all kinds of medicines, including prescription and over-the –counter

medicines. Estimated that around 230,000 are admitted in the hospital every

year because of the problems associated with the use of medicines. Examples of

this are antibiotics such as those in the sulfonamide and penicillin families

cause allergic reactions in around five per cent of the population. Skin rashes

are a common reaction.

The use of medicinal plants was mandated in the Philippines that most

of the medicinal plants in the Philippines are not scientifically proven. In 1997,
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President Fidel V. Ramos realized that the value of the traditional medicines in

the Philippines and signed into law republic Act (R.A. 8423), also known as the

traditional and alternative medicine Act (TAMA) of 1997. The law that was

created by the President created the Philippine institute of traditional and

alternative health care (PITAHC) whose mandate is to promote and advocate

the use of traditional and alternative healthcare modalities through scientific

research and product development. (Althea, 2018).

Alingatong tree is as a deciduous shrub or small tree, growing to a

height of 3 to 5 meters. Leaves are ovate to broadly elliptic-ovate, 20 to 40

centimeters long, 10 to 22 centimeters wide, entire, with shortly pointed at the

tip, and somewhat heart-shaped at the base. Upper surface is green and smooth,

the underside paler and rather densely covered with numerous, soft and stinging

hairs, abundant on the margins of the leaves. As stated in the journal of

Philippine medicinal plant the hairs of alingatong have a large bulbous base

from which projects a long tapering tube that ends in a curved tip that breaks

off easily. Contact with the leaves causes breaking of the tips of the hairs and a

poison is released that cause’s immediate and intense skin irritation. The stings

are painful and may cause the formation of blisters, immediate and possibly

progressing to confluence. With these characteristics the alingatong trees don’t

have market values and often cut by the farmers when seen because of its

harmful effect when in contact with your skin. But alingatong tree was known

to be a medicinal plant in a lot of areas in the Mindanao especially in the Santa

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Maria, Prosperidad, Agusan Del Sur one of the barangays of Agusan Del Sur to

have the practice of used alingatong (laportea batanendsis) as a medicinal tree

cure for wounds.

By these problems of the people lack of medicinal need and help, the

researchers conduct a research about the alingatong plant, to help the people in

the remote areas know the value of this plant, with no economic values of this

plant, free and can be found in their areas it is a great advantage by this far land

people to have more knowledge and experiments on these plant for future uses.

The researchers conducted a study about the chemical components of the

Alingatong Leaf Extract (ALE) and to know the coagulation time on the blood

of its extract. The researchers used phytochemical experiment to know the

components of ALE. Hence, this study, the researchers will conduct and

experiment on the blood and get the chemical properties of the Alingatong

Extract with the use of Phytochemical Screening.

1.2. Theoretical/Conceptual Framework

Figure 1.1 the official journal of Ostomy wound management stated that

Based on oriental medicine (OM) theory, wounds are considered acute or

chronic. Acute wounds often are caused by external factors, including six

external pathogenic factors (wind, cold, heat, dampness, dryness, and fire),

micro-organism invasion, and trauma. Other theories also have been gathered

by the researchers to support the idea of using the Alingatong leaf extract as an

indigenous herbal medicine. The use of herbal medicine for acute wounds was
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stated on the theory of OM that external therapy consists of various applications

of herbal plaster or herbal solutions for debridement and promoting granulation

tissue growth. Using the extract of various herbal plants for healing wounds

was one way to fasten the coagulation on the blood and heal the wound.

The use of herbal extract was stated in the research titled Oriental

Medicine and Chronic Wound Care: Theory, Practice, and Research it said that

herbal formulas and individual herbs seem effective for the treatment of chronic

wounds and acute wounds. Other theories also support the use of herbal plants

as a healing factor for wounds stated in the steady health journal 2005 the

theory of herbalism states that certain herbs possesses actual medicinal value

and can be beneficial to human health. Some of the herbs can alleviate the

symptoms of various diseases, or even cure them, while the others help to

prevent different ailments.

Hence, in this study ALE will be characterized by its chemical

composition for its medicinal value. As what the researchers have gathered in

the barangay of Santa Maria most of the residents know the plant as a plant that

were used as a wound healing plant specially in the remote areas. The chemical

composition of ALE may open a new knowledge that can be develop and can

be introduces its other beneficial uses on health. However the researchers will

only focus on its chemical composition by using the result of the phytochemical

screening and to know the duration of coagulation in the blood by using blood

test.
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Alingatong Leaf Extract (ALE)

 Chemical Composition
 Time Duration in coagulating blood

Figure 1.1

1.3. Statement of the Problem

This study is conducted to characterize alingatong leaf extract (ALE)

through phytochemical analysis and conduct an exprirement to determine the

duration of coagulation in the blood,

This specially, aims to answer the following questions:

1. Do people use Alingatong as indigenous herbal plant used for healing

wounds?

2. What are the chemical properties of Alingatong leaf extract (ALE)?

3. How long will the blood coagulate using ALE?

1.4. Significance of the Study

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The study was conducted to add more knowledge in the field of

medicine and give more emphasis on the use of herbal plants. The researchers

deemed this study significant to the:

Native people. Add more information that the alingatong leaf was a

good wound healing plant and can be used as remedies for wound instead of

buying costly medicines.

Herbal therapist. A new knowledge on curing wounds will be used in

the field of herbal medicine.

Farmers. To give more importance on the alingatong plant and be

known to more places as an herbal plant that need to be protected.

Students. Those students who don’t know the background of the ALE

wanted to have knowledge about the plant. And be able to continue the study

about the “Alingatong” plant to widen the range of knowledge about this plant.

Medicinal researchers. To give the more ideas and information on

conducting more experiments on the plant to develop new medicines.

Community. To give more information’s, ideas, and choices that there

are a lot of herbal medicine presents in our surrounding.

1.5. Scope and Limitations of the Study

The study is focused only in a blood coagulation property which is

observed through a blood sample and a drop of ALE (Alingatong Leaf Extract).

It will be witnessed through its duration on how long did it coagulated in a

blood sample.
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Moreover, there are cases that can used in some ways such as in curing

illnesses for example is anti-fungal and cures prostate cancer, but in this sort of

study we focuses only on the coagulation on the blood and by identifying its

time duration in the certain surveillance. It enables to categorizes the result of

the experiment conducted between the ALE and the blood sample and can be

the key to promote this study.

Thus, the researchers is only tasked to recognize the chemical properties

of ALE and the time occurred by the coagulation of the blood sampled in our

type of tryout which will be the main highlights of this study in order to be

valid and reliable.

In this study there are sort of terms we used and the words you may

encountered but are not familiar. Here are there definitions or details in order

for you to relate and understand the keywords and the acronyms that will

appear in this sort of research.

1.6. Definition of Terms

ALE. Alingatong Leaf Extract a plant that has hollow stinging hairs

called trichomes on the leaves, which act like hypodermic needles, injecting

histamine and other chemicals that produce a stinging sensation when contacted

by humans and other animals. Also the subject of this study.

Bulbous. Big and round often in an unattractive way having a bulb used

to describe the alingatong plant or the dependent variable of this research.

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Chemical composition. Also known as chemical makeup or simply,

composition is a concept in chemistry that has different, but similar, meanings

if referred to a single pure substance or a mixture.

Coagulation. (Widely known as clotting) is the process by which blood

changes from a liquid to a gel, forming a blood clot.

Phytochemical Screening. A test using different methods in determining

the presence and absence of phytochemicals in the sample. The extract of ALE

will undergo Phytochemical Screening in DOST-Ampayon. The results are

analyzed according to the mechanism of action a phytochemical exhibits.

Phytochemicals. Naturally occurring compounds in plants that are

responsible for the taste, color and uses of ALE. Phytochemicals present in

ALE was determined.

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CHAPTER 2

REVIEW ON RELATED LITERATURE

2.1 Conceptual Literature

Studies shows that about 25% of drugs prescribed worldwide are

derived from plants. The researchers focus on the study of the AL who is

known to be a poisonous plant. It is native to Europe, Asia, northern Africa, and

western North America, and introduced elsewhere. The species is divided into

six subspecies, five of which have many hollow stinging hairs called trichomes

on the leaves and stems, which act like hypodermic needles, injecting histamine

and other chemicals that produce a stinging sensation when contacted by

humans and other animals. The plant has a long history of use as a source of

medicine, food, and fiber.

It has been used for hundreds of years to treat painful muscles and

joints, eczema, arthritis, gout, and anemia. Today, many people use it to treat

urinary problems during the early stages of an enlarged prostate (called benign

prostatic hyperplasia or BPH). It is also used for urinary tract infections, hay

fever (allergic rhinitis), or in compresses or creams for treating joint pain,

sprains and strains, tendonitis, and insect bites.

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Although AL contains impressive amounts of vitamins and minerals,

some of the vitamins it contains include vitamins A, C, D, E, and K. It also

contains amino acids and antioxidants, which help in fighting off free radicals.

It helps detoxify the body; a diuretic, which means that it, helps in flushing out

harmful chemicals and excess liquids from the body. Herbalists have prescribed

the use of AL in treating urinary tract infections as well, because of its ability to

cleanse and dispel toxins while physical contact with the AL can cause allergic

reactions, it can be used to aid in the prevention of rhinitis, or the inflammation

of the mucous membrane in the nose.

Therefore, AL can also have bad effects regarding to individuals

allergic reaction upon contact. However, the root of it is a common treatment in

Europe for symptoms of BPH. This is a noncancerous condition that causes the

prostate gland to enlarge, making urination difficult. Studies show that stinging

nettle root may be helpful in treating this problem.

Moreover, it is needed to show that the treatment is indeed effective. It

is also said to be a cure for Eczema a dry, itchy rash that can last on sufferers

for a very long time. Because of stinging nettle’s antihistamine and anti-

inflammatory qualities, it can be a natural treatment for eczema, as the Penn

State University College of Medicine study referenced above notes.

Moreover ALE is also now in many areas for it is a stimulant and

rubefacient substance, making it very effective against various inflammatory

conditions, such as arthritis or chronic muscle pain. Research has shown that
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stinging nettle tea or herbal supplementation can effectively treat gout, soothe

muscle pain, and reduce symptoms of arthritis.

Certain products containing stinging nettle have shown, when applied to

the skin, can reduce bleeding during surgery. The product, called Ankaferd

blood stopper, is made up of alpinia, licorice, thyme, common grape vine and

stinging nettle, and has also shown evidence of reducing bleeding after dental

surgery.

The ALE was used in most places in Agusan Del Sur as a wound

healing plant. Not all people now about this except for those Lumad that was

leaving in rural areas. They have been using this plant for curing stomach ache,

fever, wound, and cough. They can use the dried or powdered leaf to stop

bleeding, pounded roots applied externally for scrofula and sometime the

decoction used for nosebleeds.

Moreover, these examples are the gathered articles of the researchers

that prove that this plant has been constantly used in rural areas as an herbal

plant. That is why the researchers want to find out if the AL has a factor of

coagulation.

A case study of Winston J Craig states that in different herbs, a wide

variety of active phytochemicals, including the flavonoids, terpenoids, lignans,

sulfides, polyphenolics, carotenoids, coumarins, saponins, plant sterols,

curcumins, and phthalides have been identifiedon various herbs that possess

hypolipidemic, antiplatelet, antitumor, or immune-stimulating properties that

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may be useful adjuncts in helping reduce the risk of cardiovascular disease and

cancer. These related case studies are based on the advantage and dis advantage

that can affect the result of our study about the factors that affect the

coagulation of the blood using ALE.

The following researches are arranged according to the chemical

composition conducted in the phytochemical results:

In relation to this case study there is an advantage of alkaloids in

tobacco leaf nicotine affected the clot-formation property of the enzyme,

thrombin, on the substrate, plasma or fibrinogen (thrombin time). From this

experimental evidence conducted by Martine Michael Ph.D., it is suggested that

nicotine does alter the clot-forming properties of thrombin on fibrinogen. Since

nicotine is present in alkaloid researchers presume that same nicotine contains

in the ALE.

Although some case studies suggests that alkaloid as antiplatelet agents

with a possible mechanism of action based on the literature in recent decade.

Bin Nan Wu Ph.D.) The research analyzed that it is a potent agent with a

description of structural relationship activity and possible lead compounds for

future drug discovery.

Thereof, in other tannins case studies it stated that it has factor in

fastening the coagulation clumps of the blood in an abstract that examined the

antibacterial action of several tannins on plasma coagulation by

Staphylococcuse (bacteria). Also in some related abstract concluded that

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tannins have also been reported exert other physiological effects, such as to

accelerate blood clotting, reduce blood pressure, decrease the serum lipid level,

produce liver necrosis, and modulate immunoresponses. Tannins and Human

Health: A Review | Request PDF. Available from: Tannins and Human Health

A Review [accessed Mar 09 2018].

Hisanori research team (KazuyasuFujii, Osamu Yamasaki,

TakashiOono, and KeijiIwatsuki) suggest that inhibition of plasma coagulation

by tannic acid is due to a decrease in the concentration of ionic calcium,

inhibition of enzyme production and hindrance of the enzyme reaction. The

mechanism of inhibition of plasma coagulation by tannic acid should be

investigated further.

Moreover in flavonoids related abstract about polyphenol compounds

belonging to flavonoids inhibits activity coagulation factor, states that blood

coagulation consists of series of zymogens which can be converted by limited

proteolysis to active enzymes leading to the generation of thrombin and

conversion of fibrinogen into fibrin by this enzyme. They concluded that they

observed only four polyphenols belonging to flavonoids group according to

Bijak M, Ponczek MB, and Nowak P. The results presented here showed that

flavonoids might be potential structural bases for design of new nature-based,

safe, orally bioavailable direct inhibitors

Moreover, an abstracts in saponnins by PubMed, that the study supports

that sapponins investigated the effects of total ginseng saponinon the cutaneous
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wound healing process using histological analysis. PubMed stated Saponins are

compounds extensively found in most plants which exist in a variety of types

and are classified based on their internal structure. Researches had revealed that

saponins can accelerate numerous biological activities including hemolytic,

anti-bacterial, anti-viral, and anti-oxidative functions. In addition, saponins

reportedly have anti-inflammatory activity which can reduce edema and skin

inflammation.

Therefore, sapponins consist with great potential in coagulation there

are some studies propose that too much sapponins could cause prolonged the

bleeding time and the whole blood decalcified clotting time, but not the plasma

decalcified clotting time.

Using Yong Nam Han, Jae-Ihn Song, and In Kyung Rhee statement

that the plasma will not decalcified clotting time. It did not affect the recalcified

clotting times of platelet-rich plasma and platelet-poor plasma while in the

presence of tissue factor the compound prolonged the reduced prothrombin

times PRP and PPP in the dose-dependent manner.

Moreover, Triterpenoids in inhibitory Effect of Triterpenoids from

Panax ginseng on blood clotting time Factor a case study about the panax

ginseng that propose that enzyme involved in the coagulation process have

received attention among attention as potential targets for the development of

oral anti-coagulants according to Schrödinger.

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Moreover, there some disadvantage of herbal remedies is used to treat a

large variety of diseases, including blood-related disorders. However, a number

of herbal preparations have been reported to cause variations in clotting time,

this is mainly by disruption of the coagulation cascade.

The compilations of plants phytochemicals investigated for effects on

the coagulation cascade. Information was withdrawn from Google Scholar, the

journal databases Scopus and PubMed. Sixty-five herbal remedies were

identified with antiplatelet, anticoagulant, or coagulating ability. Bioactive

compounds included polyphenols, taxanes, coumarins, saponins, fucoidans, and

polysaccharides.

Although research has been conducted on the effect of herbal

remedies on blood, most information relies on in vitro assays. Contradictory

evidence is present on bleeding risks with herbal uses, though herb-drug

interactions pose a threat. The researchers conclude that as the safety of many

herbals has not been proven, nor their effect on blood parameters determined,

the use of herbal preparations before undergoing any surgical procedure should

discontinued.

2.2 Research Literature

In this part contains the related studies and works of other people in

which is used to relate our sort of research. It somehow determine our research

on where it belong or categorized as sort of review or reading that may transmit

to our type of study. It also contains facts that can be added throughout our
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research. Such as promoting the coagulation effect using medicinal plants

through its extracts and chemical properties.

There was trial conducted to investigate the influence of Ginkgo biloba

special extract EGb 761 on hemostasiological parameters. There are Fifty

healthy, male volunteers underwent 7 days of crossover treatment with 2 × 120

mg/day EGb 761 and placebo in randomized sequence. Between the two

treatment phases, a washout-period of at least 3 weeks was inserted. The study's

main outcome measures were bleeding time, coagulation parameters, platelet

activity in response to various agonists and platelet morphology. (Köhler,et. al.)

Moreover, Treatment safety was investigated by clinical laboratory and

vital signs assessment and by adverse events monitoring. Among the 29

coagulation and bleeding parameters assessed, none showed any evidence of an

inhibition of blood coagulation and platelet aggregation through EGb 761.It

increase of gastrointestinal complaints during administration of the herbal

extract. (Köhler,et. al.)

Furthermore another study was established in Ankaferd Blood Stopper

(ABS), a standardized mixture of five plants, has been used historically as a

haemostatic agent but its mechanism of action remains unknown. This study

investigated the in vitro effects of ABS on haemostatic parameters. The levels

of coagulation factors II, V, VII, VIII, IX, X, XI, and XIII were not affected by

ABS. ABS has the therapeutic potential to be used for the management of
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haemorrhage and this agent should be investigated further in clinical trials.

(Goker H.et. al.)

There are also another studies which herbal plants had been observed

such as the effects of guava leaf extract on the bleeding time and the three main

mechanisms of haemostasis: vasoconstriction, platelet aggregation and blood

coagulation, were investigated. The water extract of guava leaves did not

shorten bleeding times in rats. Guava leaf extract potentiated the vascular

muscle contraction induced in rabbits by phenylephrine, and when given alone

it stimulated human platelet aggregation in vitro in a dose‐dependent manner.

On the other hand, it significantly prolonged blood coagulation; activated

partial thromboplastin time the higher the concentration of the extract, the

longer APTT was observed. Thus, a water extract of guava leaves showed

ambiguous effects on the haemostatic system. Guava leaf extract did not affect

bleeding times, it stimulated vasoconstriction and platelet aggregation but it

inhibited blood coagulation. Therefore, guava leaf extract is not recommended

as a haemostatic agent. (John Wiley & Sons, 2001)

Also, a native to Africa, is also known as “lily of the desert” or the plant

of immortality. Its name was derived from the alloeh meaning “bitter” because

of the bitter liquid found in its leaves. Egyptians recorded use of this herbal

plant in treating burns, infections and parasites as early as 1500 B.C... Its clear

gel has a dramatic ability to heal wounds, ulcers and burns by forming a

protective coating on the affected areas and speeding up the healing process.
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The fresh plant contains 96% of water and rest is essential oil, amino acids,

minerals, vitamins, enzymes and glycoproteins. Various constituents of Aloe

vera have been shown to have anti-inflammatory activity. They also stimulate

wound healing. Some clinical reports suggest topical Aloe vera gel is useful in

healing minor burns and that such application of the gel is harmless as

hypersensitive reactions to it are rare. However, in some severe burns, aloe gel

may actually impede healing (Schmidt & Greenspoon, 2008).

There are also certain plants that can heal wounds such as Azardica

indica it is commonly called as Neem and the plant has diverse medicinal

properties. Neem oil contains margosic acid, glycerides of fatty acids, butyric

acid and trace of valeric acid. Various active principles are nimbidin, nimbidal,

azardirachtin, nimbin, azadirine, gedunin, salanin. They have diverse medicinal

activities.

Neem oil is also for curing skin ailments. Oil is used for dressing for

foul ulcers, eczema and skin diseases like ringworm, scabies and mange in

dogs. It is a powerful insect repellant, anti-bacterial, anti-fungal, anti-viral, anti-

inflammatory and also strengthens the body’s overall immune responses. Neem

oil contains fatty acids which build collagen, promote wound healing and

maintain the skin’s elasticity. The active ingredients of neem oil help in the

process of wound healing and the skin is able to retain its suppleness as the

wounds heal. Neem oil has a high content of essential fatty acids. They keep the

site moist and give a soft texture to the skin during the healing process.
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Alcoholic extract of neem is useful in eczema, ringworm and scabies. Neem

leaf extracts and oil from seeds has proven anti-microbial effect. This keeps any

wound or lesion free from secondary infections by microorganisms. Clinical

studies have also revealed that neem inhibits inflammation as effectively as

cortisone acetate, this effect further accelerates wound healing.(Rajinder

Raina,2008)

St. John’s wort is a bushy perennial plant with numerous yellow

flowers. It is native to many parts of the world including Europe and the United

States. It has an age old history of safe and effective usage in many folk and

herbal remedies. It is claimed to be useful in mental depression, anxiety, sleep

disorders, menstrual cramping, sciatica and arthritis. The blossoms have been

used in folk medicine to relieve patients suffering from ulcers, gastritis,

diarrhea and nausea. This plant has an antiseptic action, relieves inflammation

and promotes healing when used externally on cut surfaces of the body. The

tincture of Hypericum spp. when given orally has a remarkable effect in

lacerated and suppurated wounds with restoration of tissue vitality (Fayazuddin,

1981). Pro-healing action of Hypericum spp. tincture is evidenced by enhanced

epithelization phase with an increase in wound contraction rate and granulation

tissue breaking strengths (Rao et al, 2008).

Tridax procumbens plant is another a native of tropical America and

naturalized in tropical Africa, Australia and Asia including India. Which is also

studied Leaf of Tridax procumbens mainly contains crude protein (26%), crude
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fiber (17%), and soluble carbohydrate (39%0 and calcium oxide (5%) (Chadha,

2007). The juice of the leaves of this plant is used by villagers to arrest bleeding

from cuts and bruises in animals. This juice accelerates two phases of healing

namely epithelization and collagenization; however it retards scar formation

and granulation (Diwan et al, 1983).

Tridax procumbens antagonized anti-epithelization and tensile strength

depressing effect of dexamethasone without affecting its anti-contraction and

anti-granulation action. (Diwan et al, 2004). The effect of various extracts

(whole plant extract, aqueous extract, butanol extract and ether fraction) of this

plant has been studied in dead space wound models. Compared to various

extracts, the whole plant extract has the greatest pro-healing activity with

increase in tensile strength and lysyl oxidase activity. Aqueous extract was also

effective in increasing lysyl oxidase but to a lesser degree (Udupa et al, 2010).

Leaf extracts of this plant also promote wound healing in both normal and

immuno-compromised (steroid treated) rats in dead space wound model. The

plant increased not only lysyl oxidase but also, protein and nucleic acid content

in the granulation tissue, probably as a result of increase in glycosamino glycan

content (Udupa et al, 2011).

However, Chromolaena odorata was first identified in Central America

and Vietnam. The aqueous extract and the decoction from leaves of this plant

have been used throughout Vietnam for the treatment of soft tissue wounds and

burn wounds. Aqueous extracts of Chromolaena odorata enhances hemostatic

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activity (Akah, 2008) and stimulates granulation tissue and re-epithelization

processes (Lee, 2009). The extract also inhibits wound contraction reversibly.

Therefore, the plant can be of much therapeutic value in minimizing post burn

scar contracture and deformities.

Hence, the oil of Hydnocarpus spp. has been used for several years as

anti-leprosy drug and as an anti-parasitic drug in the treatment of guinea worm

infestation. The oil of Hydnocarpus spp. when given orally or administered

topically helped to heal the wounds and gangrene faster in leprosy and diabetic

patients (Manjrekar, 2009). The wound healing effect of oil of Hydnocarpus

spp. was studied with reference to collagenation and the strength of scar tissue

(Oomen et al, 2000). The drug treated group showed a significant increase in

strength of scar tissues in the incision wound model and also increased the

strength of collagen tissue and hydroxyl-proline content in the dead space

wound model. Hydnocarpus oil administered orally promoted epithelization,

but not wound contraction (Oomen et al, 2000). External application of oil of

Hydnocarpus spp. and its paste significantly shortened the epithelization period

when compared to control group. Oil may act as adjuvant in healing of wounds

and ulcer in leprosy patients and therefore, may be clinically useful.

Another plant is Helianthus annus Linn an ornamental annual herb, with

erect, rough and hairy stem is common in Indian Gardens in swampy areas. In

traditional medicine the plant is used by tribals for inflammation of eyes, sores,

dysuria, colic, tiger bites and bone fractures (Jain & Tarafdar, 2001). In a study
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the alcoholic extract of whole plant of H. annus applied in the form of an

ointment on the excised wound of rat led to a significant reduction in total

healing period. This has been confirmed by histology where earlier appearances

of fibroblasts were seen. Early appearance and higher accumulation of muco-

polysaccharides has been stated as indicators of hastened repair (Deshpande et

al, 2009).

Though Urcuma longa Linn. Commonly known as turmeric and haldi in

Hindi. C. longa has been reported to possess anti-bacterial, anti-fungal and anti-

inflammatory activities (Chopra et al,). The part used are rhizomes and it

contains curumin (diferuloyl methane), turmeric oil or turmerol and 1,7-bis, 6-

hepta-diene-3, 5-dione. Curcumin has potent anti-inflammatory and analgesic

activities (Srimal et al, 2011).

Likewise Volatile oil isolated from C. longa also exhibits antibacterial

and potent anti-inflammatory activity. Curcuma longa also contains protein,

fats, vitamins (A, B, C etc) all of which have an important role in would healing

and regeneration. Turmeric has been used for treating the wounds in the rats

(Rao et al, 2003). The anti-inflammatory property and the presence of vitamin

A & proteins in turmeric result in the early synthesis of collagen fibers by

mimicking fibroblastic activity (Kumar et al, 2008). Juice of the fresh rhizome

is commonly applied to recent wounds, bruises & leech bites. A paste of

turmeric & leaves of Justica adhatoda with cow urine is rubbed on skin affected
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with prurigo & eczema. It can also be mixed with ginger oil to prevent skin

eruptions.

Additionally Centella asiatica (Brahmi) also known as “gotu kola”, is

the main herb in Ayurveda for nervous system, it is used in the repair of

nervous tissue from crushing trauma, such as spinal injury, neuromuscular

disorders, and to increase general brain function and memory concentration. It

is used extensively in the treatment of leprosy, a host of skin conditions

including cellulites, varicose vein and wounds. The active principles of Centella

asiatica are triterpenes and asiaticoside which are responsible for promotion of

rapid wound healing (Shukla et al, 2008). Aqueous extract of Centella asiatica

suspended in 5% propylene glycol promoted wound healing on topical

administration in experimentally induced open wounds in rats as compared to

other extracts (alcoholic, petroleum ether and chloroform). This was evidenced

by the increase in collagen content and thickness of epithelium (Rao et al,

2006). However, Suguna et al, (2008) demonstrated that alcoholic extract of

Centella asiatica orally or topically improved the rate of wound healing in rats.

Topical administration of the aqueous extract increased cellular proliferation,

promoted the collagen synthesis at the wound site as evidenced by the increase

in DNA, protein, collagen content of granulation tissue and in tensile strength.

The treated wound epithelized faster as compared to control. Among the

various formulations (ointment, cream and gel) of aqueous extract, the process

of healing was better with gel formulation.

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Last of all is the healing effects of Ocimum sanctum, and Begia odorata

on infected experimental wounds in laboratory animals have been reported. All

these plants, notably Ocimum sanctum promote healing (Thaker & Anjaria

2006). The aqueous extract Euphoribia nerrifolia when applied topically

facilitates the healing of surgically produced cutaneous wounds in guinea pigs.

It increases the gain in tensile strength, DNA content and promoted

epithelization (Rasik et al, 2006). Alcoholic extract of Indigofera aspalathoides

has analgesic, anti-inflammatory and wound healing effect. The crude betal nut

extract and its polyphenols promoted healing of incision and dead space

wounds (Padmaja et al, 2008). Fresh leaves of Kalanchoea integra showed

encouraging results in healing inflammatory conditions associated with

wounds. Mango butter which is extracted from the seeds of Mangifera indica is

known to have wound healing properties.. Anecdotal evidence, some clinical

observations, animal model studies and few randomized clinical trials support

the efficacy of honey in managing wounds (Molan, 2000).

Thereof, in other tannins case studies it stated that it has factor in

fastening the coagulation clumps of the blood in an abstract that examined the

antibacterial action of several tannins on plasma coagulation by

Staphylococcuse (bacteria). Also in some related abstract concluded that

tannins have also been reported exert other physiological effects, such as to

accelerate blood clotting, reduce blood pressure, decrease the serum lipid level,

produce liver necrosis, and modulate immunoresponses. Tannins and Human

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Health: A Review | Request PDF. Available from: Tannins and Human Health

A Review [accessed Mar 09 2018].

Moreover, there some disadvantage of herbal remedies is used to treat a

large variety of diseases, including blood-related disorders. However, a number

of herbal preparations have been reported to cause variations in clotting time,

this is mainly by disruption of the coagulation cascade.

Generally this related researchers approved that medicinal plants having

its chemical properties has the capability in healing wounds. Especially in

external wounds. However this related researches synchronized every medicinal

plants occurred. Suguna et al alcoholic extract of Centella asiatica orally or

topically improved the rate of wound healing. And the aqueous extract from

leaves of this plant have been used for the treatment of soft tissue wounds and

burn wounds. (Akah, 2008). These natural agents occurring from medicinal

plants induce healing and regeneration of the lost tissue by multiple

mechanisms. However, there is a need for scientific validation, standardization

and safety evaluation of plants of the traditional medicine before these could be

recommended for healing of the wounds. ( Rajinder Raina,et al)

CHAPTER 3

RESEARCH METHODS

This chapter includes the information concerning oh how does the

experiment conducted, and how does the data gathered and analyze.

3.1 Research Design

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This study follows the experimental method type of research. Its main

topic is to determine if the ALE will have factors that can help coagulation in

which concerning only in external wounds.

3.2 Setting of the Study

This study had revolved on the knowledge about the evident practice of

using ALE as an herbal remedies and the conduct of tests to further determine

its coagulation factors in wounds and cuts. An interview is conducted at Santa

Maria, Prosperidad, Agusan Del Sur for documentation.

Furthermore, the leaf is collected from Santa Maria, Prosperidad,

Agusan Del Sur where the practice of using the leaf extract for wounds and cuts

is mostly used. The leaf was ensured that no other insects and dirt will

contaminate the extract.

The ALE will then be transported to the laboratory- DOST, Ampayon to

undergo phytochemical screening

3.3 Data Gathering Procedure

The leaf of alingatong was gathered and cut in the branch for the leaf of

alingatong has stinging hairs that can be so irritated when in contact in the skin.

The leaf was cut fresh in the branch of the Alingatong tree. The leaf that will be

gathered should make a 100 mL extract as a requirement for the Phytochemical

Screening. Figure 3.2 (a) shows the exact alingatong tree where the alingatong

leaf (Figure 3.2 (b)) were cut.

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Figure 3.2 (a) Alingatong tree Figure 3.2 (b) Alingatong Leaf

The leaf had been cut into small pieces, was labeled and then placed in a

sealed plastic bag. The leaves were ensured that no other dirt, water, and other

chemicals may contaminate the leaf. . This was transported right after it was

gathered to the testing facility at Department of Science and Technology -

Ampayon, Agusan Del Sur situated at Caraga State University.

It had undergone extraction procedures. The extraction method used

was by a rotary evaporator. The extract was then subjected to phytochemical

screening following the procedures from the book entitled, “A Guidebook to

Plant Screening: Phytochemical and Biological” to determine the

phytochemicals present in the sample using many test to determine the

chemical present in the ALE. Figure 3.3 shows the deferent Test tube test to

determine the presence of these components. In the Alkaloids test a

confirmatory test and quaternary bases and Amino Oxide test was conducted, in

the steroid test a Keller-Killiani test and Liebermann-burchard test was

conducted, in the test for flavonoids a Bate-Smith and Metcalf method was

conducted, for the test of saponins a Froth test was conducted, and the last for

tannin test a Ferric Chloride test was conducted.

Test Chemicals Laboratory Test Tube Method

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Confirmatory Test

Test for Alkaloids Test for Quaternary bases and /

or Amino Oxide

Keller-Killiani Test

Test for Steroids Liebermann-Burchard test

Test for Flavonoids Bate-Smith and Metcalf Method

Test for Saponins Froth test

Test for Tannins Ferric Chloride Test

Figure 3.3 Phytochemical Screening Test

After the Phytochemical Screening, the results were given after fourteen

(14) working days given by the condition of the testing facility. The researchers

were then conducted the blood test to determine the duration of coagulation in

the blood. The blood was classified in three type’s blood type O, A, A/B and B,

each of the blood type was given three samples to get the mean and compare

which of the three types was the fastest to coagulate using ALE. Each the three

blood samples will be compared by its normal coagulation and the coagulation

using the ALE. The method we will used is “Slide Method or Drop Method”.

“Slide method or drop method”

Materials

 70% alcohol

 Sterile gauze

 Sterile disposable lancet

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 Stopwatch

 Clean glass slide

 Needle

Procedure

1. Disinfect the site of puncture with 70% of alcohol

2. Puncture to depth of 3mm

3. Wipe off the drop of blood

4. Start the timer soon as the 2nd drop of blood appears

5. Transfer to a clean glass slide three separate drops of blood

6. Observe for clot formation or coagulation by using the lancet to check for

fibrin threads in the blood. This appear as a “threadlike” substance that clings

to the tip of the lancet

7. Record the time when the first thread was formed

Normal time

 2-4 minutes

3.4 Data Analysis

Blood NC AC NC AC NC AC Mean Mean

Type 1st 1st 2nd 2nd 3rd 3rd NC AC
attempt attempt attempt attempt attempt Attemp
t
AB 99s 98s 96s 96s 101s 98s 98.67s 97.3s
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A 158s 134s 156s 132s 157s 132s 157s 132.7s

O 113s 101s 110s 98s 125s 103s 116s 100.7s

B 74s 68s 78s 74s 83s 68s 78.3s 70s

Legend: NC- Normal Coagulation

AC- ALE Coagulation

Figure 3.4 Blood Coagulation duration result using the Slide method.

Figure 3.4 shows that there is a possibly that Alingatong Leaf Extract (ALE)

can make the coagulation faster compared to its normal coagulation in the

four blood types, Hence the data we gathered is from the method we user

widely known as Sliding Method or the Drop-off Method. This data will be

the main highlights of our research, so that our study can be successful,

moreover the researchers used a lot of Laboratory apparatus in conducting our

experiment. We used the four blood types of blood got by the living person’s

blood, However after we interviewed some of the individual of Brgy. Sta

Maria, Agusan Del Sur and got the result of Phytochemical analysis. The

researchers will proceed to analyze the data in the coagulation of ALE in the

blood and determining its blood duration.

CHAPTER 4

PRESENTATION, ANALYSIS, AND INTERPRETATION OF DATA

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This chapter shows how the data was presented with the use of the

results given by the other chapter, it will also show how the data was analyze

and interpreted by the researchers.

4.1. Alingatong plant used for healing wounds

Herbal medicine was mostly and widely used now a day for the

increasing price of medicinal drugs. The developments of new drugs for

treatments are also one of the priorities these days. An interview was conducted

in the place of the Santa Maria, Prosperidad Agusan Del Sur with its residents

for the documentation on the use of ALE as an herbal remedies for wounds.

Person 1 said that “Yes I used Alingatong plant in treating my muscle pain by

the used of decoction and secondhand the leaf through applying it in the part

where the pain is located”.

Person 2 stated that “Yes I used Alingatong plant in the treatment of my

arthritis by boiling it and drink the soup of the boiled plant”.

Person 3 communicated that “Yes I used Alingatong plant in threatening such

wounds by getting its leaf extract and applied it in my wounds”.

4.2. Results of the Phytochemical Screening

The researchers conducted a phytochemical screening on its chemical

components for wound healing. Table 4.1 the phytochemical result shows that

there are a lot of properties in the ALE that are present. The alkaloid in the ALE

was given a result of (++) means that it has a secondary alkaloid present in the

ALE. The steroids also have (+) in the deoxysugar and unsaturated alkaloid.
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Flavanoid also have (+) meaning that it has a presence of that chemical. The

Saponins have (+) also meaning that ALE has presence of this chemical. The

(+) also in the tannins means that it has a presence of this chemical having a

color of blue black indicates also the presence of hydrolysable tannins.

PHYTOCHEMICAL RESULT

SCREENING

Alkaloids

Confirmatory test

(+) primary Alkaloid

++
(++) secondary Alkaloid

(+++) tertiary Alkaloid

Steroids

Keller-Killiane test: For +

2-deoxysugar +

Liebermann-

burchardtest=for Unsaturated

Steroids

Flavonoids

Bate-Smith & Metcalf +

Method= For

Leucoanthocyanins
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Saponins

Froth Test +

Tannins

Ferric Chloride Test

*brownish-green color indicate the

+
presence of condensed tannins
Blue-black
*blue-black color indicate the

presence of hydrolysable tannins.

Figure 4.1 Phytochemical Screening Result

(+) indicate the presence of the chemical

(-) indicate the absence of chemical

4.3. The time duration of blood coagulation with ALE

Blood NC AC NC AC NC AC Mean Mean

Type NC AC

AB 99s 98s 96s 96s 101s 98s 98.67s 97.3s

A 158s 134s 156s 132s 157s 132s 157s 132.7s

O 113s 101s 110s 98s 125s 103s 116s 100.7s

B 74s 68s 78s 74s 83s 68s 78.3s 70s

Figure 4.2 ALE Coagulation Time Duration Result

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Figure 4.2 shows that ALE is proven and tested to fasten the coagulation of

blood by testing its time duration, Moreover the ALE was applied through the

three samples of blood which is the type A, A/B, B and O and undergoes into

three trials where in the type A blood dominates the fastest coagulation of the

three blood samples. It is proven through the comparison of its normal

coagulation and the ALE coagulation which was used by a “Sliding Method

or Drop-off Method, Hence the study validates by getting the mean of result

of its time duration between the comparison of the mean normal coagulation

and the mean ALE coagulation. We the researchers asked the permission

through our advisee to conduct the such experiment and by researching it in

the internet on how to determine the coagulation time in a blood and we

succeed in conducted the experiment by the used also of the laboratory

equipment that are found in our “Bio Chem.” Laboratory and Physics

Laboratory. The researches extracted by using the specific laboratory

equipment and by filtering it with a clean 100% cotton, the researchers also

found out that there is a chemical reaction due to its bad smell and it greenish

color. We practically used the blood from a living person. Moreover the ALE

can be used as coagulation agent in such wounds and best applied in wounds

that the blood type that is A.

CHAPTER 5

SUMMARY, CONCLUSION AND RECOMMENDATIONS

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This chapter includes the summary of the study, conclusion and

recommendations for the improvement of the study.

5.1 Summary

This study desires to determine the used Alingatong Leaf Extract

(ALE), specifically the aims to:

1. Found out the used of ALE in the residents of Sta Maria, Agusan

Del Sur as an indigenous plant in their Barangay.

2. To evaluate chemical composition of ALE by the used of

Phytochemical Analysis.

3. Observed the time duration of ALE by its coagulation when applied

in the four type of blood which are A, A/B, B, and O.

ALE as a treatment for wounds and cuts is a common practice in the

residents of Sta. Maria, Prosperidad, Agusan Del Sur for generations which

makes it a medicinal plant. ALE is a common plant and can only be found in

the deep of the jungle where in known for other human a plant with no uses and

harmful due to its stinging hairs. Where in some other native people said that it

is used for fastening the healing of wounds and cuts. The results of the

phytochemical test showed that the extract has phytochemicals present of

Alkaloids, Steroids, Flavonoids, Triterpenes, Saponins, Tannins.

Alkaloids exhibit antimalarial, Steroids exhibit antidiarrhoeal activity.

Flavonoids exhibit antimicrobial and antidiarrhoeal, Saponins exhibit

antidiarrhoeal, anticancer and anthelmintic. Tannins and Triterpenes, exhibit

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antimicrobial, antidiarrheal and anthelmintic. On the other hand, out of the five

phytochemicals tested, alkaloids are not present in the extract.

Therefore, phytochemical test on the sample determines the presence of

Alkaloids. Wherein Alkaloids Steroids, Flavonoids Saponins, Tannins are the

factors that can lead in coagulation activity on treating wound and cuts.

However, by conducting the experiment using the Sliding Method or Drop-off

Method we the researchers observed that ALE is an effective used in fastening

the coagulation time duration as it is applied in the four type of blood which are

the type A, A/B, B, and O by getting its mean through the comparison of the

normal coagulation and the coagulation of ALE.

5.2. Conclusion

Therefore Alingatong plant is used by the residence of the Brgy, Sta

Maria, Agusan Del Sur. Results of the phytochemical analysis showed that the

ALE is presented with Alkaloids Steroids, flavonoids, steroids, saponins and

tannins. This further proved that the extract has a coagulation activity by

evaluating the mechanism of action of the phytochemicals. ALE has a strong

presence of coagulation activity and was proven through the experiment using

the Sliding Method or Drop-Off Method and observed and compared its normal

coagulation and the coagulation of ALE in the four types of blood.

5.3 Recommendation
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Based on the conclusions and the summary of the findings of the study,

here are the recommendations for the improvement of the research and for

future purposes:

5.3.1. To conduct an interview it must be in all the remote areas of the

Philippines where in Alingatong plant is used.

5.3.2. To conduct the experiment in determining the blood coagulation it

must be approved by the DOH and any blood Doctors.

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Lasco, G. (2009). Lipa (tropical wood nettles): an overview.

liu, w. J. (2007). Traditional Herbal Medicine Research Methods. china:

WILEY publishers.

M.D., D. A. (n.d.). Stinging Nettle Benefits. Retrieved from

herbwisdom.com: http://www.herbwisdom.com/herb-nettle.html

Medicine, G. E. (1994). Retrieved from encyclopedia.com:

http://www.encyclopedia.com/plants-and-animals/plants/plants/nettle

Min Jin, 1. Y.-F.-Q.-P.-F. ( 2014). Effect of Toona microcarpa Harms

Leaf Extract on the Coagulation System. china.

plant, p. m. (n.d.). lipa (laportea meyeniana warb.). Retrieved from

http://www.stuartxchange.com/Lipa

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https://www.itis.gov/servlet/SingleRpt/SingleRpt?

search_topic=TSN&search_value=19152#null

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The National Center for Teacher Education

Ghorbanibirgani, A. ((2013)). The Efficacy of Stinging Nettle (Urtica

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Blind Study in 100 Patients. Iran: Iran Red Crescent Med J.

sachiehealth. (2012, APRIL 21). HERBAL REMEDIES FOR

FILIPINOS. Retrieved from Philippine Traditional and Alternative Medicine:

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alternative.html

Sanjeev Palta, R. S. ( 2014 ). Overview of the coagulation system. india:

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STURLUSON, T. (2014). History of Herbal Medicine. THE HERBAL

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APPENDIX A

Semi-structured Interview

Case Report no. 1

NAME
ADDRESS
GENDER
AGE
OCCUPATION
MEDICINAL STATUS

Have you ever used Alingatong plant as treatment which is

evident in your place?
Yes (Answer the questions below) No

1. Specify the illness/es or disease/s, any health problem, etc. that

USE OF is treated or applied with the Alingatong plant for treatment.
ALINGATON
2. What are the sensations felt by applying the extract?
G FOR
MEDICATION
3. Is Alingatong effective in the treatment?

4. Do you recommend Alingatong extract as a treatment?

5. Do you have any other recommendations on the manner of

using or applying this treatment?
 PHILIPPINE NORMAL UNIVERSITY

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Case Report no. 2

NAME
ADDRESS
GENDER
AGE
OCCUPATION
MEDICINAL STATUS

Have you ever used Alingatong plant as treatment which is

evident in your place?
Yes (Answer the questions below) No

1. Specify the illness/es or disease/s, any health problem, etc. that

USE OF is treated or applied with the Alingatong plant for treatment.
ALINGATON
2. What are the sensations felt by applying the extract?
G FOR
MEDICATION
3. Is Alingatong effective in the treatment?

4. Do you recommend Alingatong extract as a treatment?

5. Do you have any other recommendations on the manner of

using or applying this treatment?

Case Report no. 3

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NAME
ADDRESS
GENDER
AGE
OCCUPATION
MEDICINAL STATUS

Have you ever used Alingatong plant as treatment which is

evident in your place?
Yes (Answer the questions below) No

1. Specify the illness/es or disease/s, any health problem, etc. that

USE OF is treated or applied with the Alingatong plant for treatment.
ALINGATON
2. What are the sensations felt by applying the extract?
G FOR
MEDICATION
3. Is Alingatong effective in the treatment?

4. Do you recommend Alingatong extract as a treatment?

5. Do you have any other recommendations on the manner of

using or applying this treatment?

APPENDIX B
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Results of the Phytochemical Screening

APPENDIX C
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Documentations

Actual Photo in the DOST, Ampayon during the

Phytochemical Test

Actual Photo Sample ALE

Actual Photo Left Side Type B with ALE right side

ALE
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Curriculum Vitae

Personal Information

Name: Ryan Mark C. Aunzo

Birthdate: October 10, 1998

Birthplace: Salvacion, Prosperidad Agusan del sur

Address: P-2, Salvacion, Prosperidad, Agusan del sur

Civil Status: Single

Religion: Roman Catholic

Nationality: Filipino

Parent’s Name

Father: Joel M. Aunzo

Mother: Maria Lisa C. Aunzo

Academic Background

Tertiary: Philippine Normal University - Mindanao

Secondary: Agusan Sur National Science High School

Primary: Salvacion Elementary School

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Personal Information

Name: Wilven Lloyd P. Gaviola

Birthdate: March 12, 1997

Birthplace: Sta. Maria, Prosperidad, Agusan del Sur

Address: Sta. Maria, Prosperidad, Agusan del Sur

Civil Status: Single

Religion: ECOFI

Nationality: Filipino

Parent’s Name

Father: Baldwin A. Gaviola

Mother: Anecita P. Gaviola

Academic Background

Tertiary: Philippine Normal University - Mindanao

Secondary: Agusan Sur National High School

Primary: Pilot Central San Francisco Elementary School

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Personal Information

Name: Florencio C. Maceda II

Birthdate: June 30, 1999

Birthplace: Pisaan,SanFrancisco, Agusan del Sur

Address: P-3, Pisaan, SanFrancisco,, Agusan del Sur

Civil Status: Single

Religion: Roman Catholic

Nationality: Filipino

Parent’s Name

Father: Gil B.Maceda

Mother: Vita C. Maceda

Academic Background

Tertiary: Philippine Normal University - Mindanao

Secondary: Agusan Sur National High School

Primary: Pisaan Elementary School